Martel F, Azevedo I, Osswald W
Institute of Pharmacology and Therapeutics, Faculty of Medicine, Porto, Portugal.
Naunyn Schmiedebergs Arch Pharmacol. 1994 Aug;350(2):130-5. doi: 10.1007/BF00241086.
Isolated rat hepatocytes were incubated with 200 nmol/l 3H-(-)-noradrenaline or 50 nmol/l 3H-(-)-adrenaline for 15 min, in Krebs-Henseleit solution at 37 degrees C, gassed with 95% O2 5% CO2. Monoamine oxidase and catechol-O-methyl transferase were inhibited with pargyline (500 mumol/l) and Ro 01-2812 (3,5-dinitropyrocatechol; 2 mumol/l), respectively. Total radioactivity present in the cells, which corresponded mostly to intact 3H-amine, was measured. The content of 3H-noradrenaline increased with time of incubation, a plateau having been reached after 15 min of incubation. After 15 min of incubation, the cell: medium ratio for 3H-noradrenaline and 3H-adrenaline was 0.6-0.7. Desipramine (an inhibitor of the neuronal uptake of catecholamines-uptake1; 1 mumol/l) did not affect the uptake of either 3H-noradrenaline or 3H-adrenaline into hepatocytes. Corticosterone (an inhibitor of the extraneuronal uptake of catecholamines-uptake2; 40 mumol/l) slightly inhibited (by 28%) the uptake of 3H-adrenaline, and did not significantly reduce 3H-noradrenaline uptake. Probenecid (an inhibitor of the renal transport of organic anions; 100 mumol/l) did not influence the amount of either 3H-noradrenaline or 3H-adrenaline in hepatocytes. Cyanine 863 (an inhibitor of the renal transport of organic cations; 10 mumol/l) decreased by 62% the uptake of 3H-adrenaline into cells but did not significantly affect 3H-noradrenaline uptake. Bilirubin (a substrate of a hepatic transport for organic anions; 200 mumol/l) produced a significant increase (50%) in the amount of 3H-noradrenaline and 3H-adrenaline present in the cells.(ABSTRACT TRUNCATED AT 250 WORDS)
将分离的大鼠肝细胞在含有200 nmol/l 3H-(-)-去甲肾上腺素或50 nmol/l 3H-(-)-肾上腺素的Krebs-Henseleit溶液中,于37℃、95% O2和5% CO2的气体环境中孵育15分钟。分别用优降宁(500 μmol/l)和Ro 01-2812(3,5-二硝基邻苯二酚;2 μmol/l)抑制单胺氧化酶和儿茶酚-O-甲基转移酶。测量细胞中存在的总放射性,其大部分对应于完整的3H-胺。3H-去甲肾上腺素的含量随孵育时间增加,孵育15分钟后达到平台期。孵育15分钟后,3H-去甲肾上腺素和3H-肾上腺素的细胞:培养基比值为0.6 - 0.7。地昔帕明(一种儿茶酚胺神经元摄取抑制剂-摄取1;1 μmol/l)不影响3H-去甲肾上腺素或3H-肾上腺素进入肝细胞的摄取。皮质酮(一种儿茶酚胺非神经元摄取抑制剂-摄取2;40 μmol/l)轻微抑制(28%)3H-肾上腺素的摄取,且未显著降低3H-去甲肾上腺素的摄取。丙磺舒(一种有机阴离子肾转运抑制剂;100 μmol/l)不影响肝细胞中3H-去甲肾上腺素或3H-肾上腺素的量。花菁863(一种有机阳离子肾转运抑制剂;10 μmol/l)使3H-肾上腺素进入细胞的摄取减少62%,但未显著影响3H-去甲肾上腺素的摄取。胆红素(一种有机阴离子肝转运底物;200 μmol/l)使细胞中3H-去甲肾上腺素和3H-肾上腺素的量显著增加(50%)。(摘要截短至250字)